Gökce Ilayda Söztekin, MSc
Department(s): Department of Pathobiology of the Nervous System (Center for Brain Research)
Position: PHD Student
Location: Spitalgasse 4
Telephone: 40160 -34092
Human genetic studies have highlighted a link between schizophrenia and alterations in nicotinic acetylcholine receptor (nAChR) genes including CHRNA7 (coding for α7 subunit) and CHRFAM7A. CHRFAM7A occurs only in humans and encodes a partially duplicated version of the α7 nAChR subunit lacking the agonist binding site. CHRNA7 and CHRFAM7A are located in an unstable region of the genome, resulting in copy number variations (CNVs) of the two genes. In our project, our aim is to investigate the impact of dupα7 subunits on α7 receptors in human iPSC-derived neurons. This study is funded by the Austrian Science Fund (FWF: I3778).
Cell culture Cortical neurons are differentiated from human induced pluripotent stem cells (hiPSCs) with either overexpressing or lacking CHRFAM7A to mimic the copy number variations. Immunocytochemistry To characterize human iPSC- derived neurons neuron specific markers are used. Fura-2 Ca++ imaging In order to investigate the effect of somatic α7 receptors characterized by high Ca++ permeability and fast desensitization on intracellular Ca++ levels Fura-2 is used for Ca++ imaging. Fura-2 is a ratio-metric fluorescent dye which binds to free intracellular calcium. By using α7 receptor specific compounds on cell lines having different copy numbers of CHRFAM7A, intracellular Ca++ level changes can be observed. Patch Clamp Electrophysiology For functional characterization of human iPSC-derived neurons and to assess the pre-synaptic α7 receptors on a cellular level, whole- cell patch clamp is performed. By using α7 receptor specific compounds, impact of neurotransmitter release on post-synaptic cells can be investigated.
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